Quantitative study of synaptic boutons on frog intracardiac neurons

Abstract

The frog is a useful model to study the structure and function of intracardiac neurons. The goal of this study was to evaluate the size and distribution of synaptic boutons on the intracardiac neurons in the frog Rana temporaria. Interatrial septa from four animals were double-labelled immunohistochemically for the cholinergic marker choline acetyltransferase (ChAT) and the marker of synaptic vesicles synaptophysin (SYP). One hundred intracardiac neurons were analysed by confocal microscopy. Terminals of preganglionic axons were strongly positive for ChAT, while synaptic boutons were strongly positive for both ChAT and SYP. The number of synaptic boutons per neuron ranged from 2 to 121 and was 10±2 (mean±SE). The total area of synaptic boutons ranged from 6 μm 2 to 270 μm 2 and was 98±6 μm 2 . The largest total area of synaptic boutons was found on the axonal half of neuronal soma (59±4 μm 2 ). The total areas of synaptic boutons on both the non-axonal half of soma and the proximal axon were smaller (36±3 μm 2 ; 6±2 μm 2 ; P<0.001). Synaptic boutons occupied 13±1% of the area of the neuronal soma profile. Conclusions: 1) The axonal half of the soma of the frog intracardiac neuron is more densely innervated than the non-axonal half of the soma. 2) The axosomatic and axoaxonic synapses are present on frog intracardiac neurons. The study provides a framework for further experimental studies on the formation and rearrangement of synapses on frog intracardiac neurons.